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目的 :探索表达人纤溶酶原半胱氨酸卷曲区 5 (hPK 5 )的最佳菌株 ,为研究其抑制内皮细胞增殖和迁移的活性 ,开发肿瘤治疗和预防肿瘤转移的新药提供前提。方法 :在将人纤溶酶原半胱氨酸卷曲区 5 (hPK 5 )基因与原核表达载体pBV2 2 0进行体外重组获得表达质粒 pBV2 2 0 /hPK 5 ,采用氯化钙转化法将重组质粒分别导入BL2 1(DE3)、DH5α ,JM10 9和BL2 1(DE3) pLyss 4种工程菌 ,在温控诱导表达条件下进行相同表达条件和优化表达条件下的hPK 5因子表达差异性研究。 结果 :工程微生物 (本研究为工程菌株 )的筛选、培养条件的建立、外源基因表达条件的建立是优化基因工程技术体系的重要技术环节。结论 :本研究为hPK5因子的进一步深入研究以及产业化的发展奠定了基础
OBJECTIVE: To explore the optimal strain expressing human plasminogen curly 5 (hPK 5), providing the premise for the study of its activity of inhibiting the proliferation and migration of endothelial cells and developing new drugs for tumor therapy and prevention of tumor metastasis. Methods: Expression plasmid pBV2 2 0 / hPK 5 was obtained by in vitro recombination with human prokaryotic cysteine curl region 5 (hPK 5) gene and prokaryotic expression vector pBV220. The recombinant plasmids were transformed by calcium chloride Four kinds of engineering bacteria, BL2 1 (DE3), DH5α, JM109 and BL2 1 (DE3) pLyss, were respectively introduced into E.coli BL21 (DE3) pLyss to study the difference of expression of hPK 5 under the same expression conditions and under the condition of temperature-induced expression. Results: The screening of engineering microorganisms (engineering strain), the establishment of culture conditions and the establishment of exogenous gene expression conditions are the important technical links in the optimization of genetic engineering technology system. Conclusion: This study laid the foundation for further in-depth study of hPK5 factor and industrialization